The Cell Microinjection Shared Resource provides a state-of-the-art cell microinjection facility for Cancer Center investigators engaged in peer-reviewed grant-funded research or studies that will lead to the submission of grant proposals. This facility allows investigators at St. Jude Children's Research Hospital to incorporate microinjection into experiments designed to analyze proteins and DNA. Microinjection offers several advantages over conventional techniques but the paramount one is the ability to precisely control the amount and timing of material to be introduced directly into cells, where their biological effect can be directly observed. The facility also has the capacity to directly image living cells in real time. Assistance with analysis of the injected cells is also provided. Standard methodologies for analysis included time-lapse photography, BrdU labeling and mitotic index quantitation for cell cycle alteration and Annexin V, TUNEL, caspase activation, Mitotracker, JC-1 and rhodamine 123 for examination of apoptotic events. A full-time technician staffs the cell microinjection Shared Resource, located in the Thomas Tower DP008. This Shared Resource is equipped with a Zeiss Axiovert microscope with an automatic stage controller and heated stage. An Eppendorf micromanipulator and microprocessor-controlled semi-automatic injector (Transjector), a tissue culture incubator, three cameras, including a CCD camera for documenting results and two computers containing all common Microsoft programs and Metamorph and Axiovision imaging software. The projected total budget for this Shared Resource in year -25 of this grant is $86,156, of which 70% ($60,977) is requested from the CCSG; the remainder of the budget (30%; $25,179) will be provided by SJCRH institutional funds. Essentially 99% of the usage of this Shared Resource is by Cancer Center members for peer reviewed funded projects.